Microfibril-associated glycoproteins attenuating pulmonary fibrosis

微纤维相关糖蛋白减轻肺纤维化

• 批准号: 10595656
• 负责人: Jeffrey Koenitzer
• 金额: $ 15.62万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10595656
• 关键词: Acceleration; Advisory Committees; Affinity; Attenuated; Binding; Bioinformatics; Biological Assay; Biology; Bleomycin; Bone Marrow; Cell Nucleus; Cells; Characteristics; Chromatin; Collaborations; Compensation; Data; Deposition; Development; Disease; Disease Progression; Exhibits; Extracellular Matrix; Family member; Fibroblasts; Fibrosis; Follow-Up Studies; Gene Expression; Gene Expression Regulation; Genes; Glycoproteins; Goals; Growth Factor; Homeostasis; Human; Hypoxia; Image; Immune; Immune response; In Situ; Individual; Inflammatory; Inflammatory Response Pathway; Injury; Interstitial Lung Diseases; Knock-out; Knockout Mice; Lung; Lung Transplantation; Lung diseases; MFAP1 gene; Macrophage; Mediating; Mentors; Mesenchymal; Microfibrils; Modeling; Mus; Names; Pathogenesis; Pathway interactions; Phase; Phenotype; Physicians; Play; Population; Process; Production; Proliferating; Protein Secretion; Proteins; Pulmonary Fibrosis; Recovery; Regulation; Reporting; Research Personnel; Role; Scientist; Signal Transduction; Specificity; Supportive care; Tissues; Training; Transforming Growth Factor beta; Transposase; Up-Regulation; Work; airway obstruction; antifibrotic treatment; career; career development; cytokine; epigenome; epigenomics; experimental study; fibrogenesis; fibrotic lung; idiopathic pulmonary fibrosis; immune cell infiltrate; immune function; improved; in vivo; inhibitor; interest; lung histology; lung injury; mortality; novel strategies; receptor; response; side effect; single cell sequencing; single nucleus RNA-sequencing; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transcriptomics; transforming growth factor beta3

Project Summary/Abstract Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with limited treatment options and unclear pathogenesis. Extracellular matrix (ECM) deposition by proliferating fibroblast populations drives the progressive airflow limitation and hypoxia characteristic of IPF, mediated by the cytokine transforming growth factor beta (TGFB). Direct inhibition of TGFB leads to intolerable side effects, and there is increased interest in therapies that instead modulate its regulation. Microfibril-associated glycoproteins 1 and 2 (Magp1 and Magp2, or Magps) are fibroblast-produced proteins which are secreted to ECM, anchor to microfibrils, and bind TGFB to limit its signaling under basal conditions, but have no known role in fibrosis. Magp knockout (KO) mice exhibit increased TGFB signaling in various tissues and have immune cell alterations due to bone marrow TGFB effects. In a single cell RNASeq screen, the genes encoding these proteins were markedly upregulated in fibroblasts in fibrotic lungs, and combined KO of Magp1 and 2 led to more severe fibrosis after bleomycin injury in mice. This suggests that Magps act as inhibitors of fibrotic signaling and follow up studies in Magp1 and 2 individual KO mice show that deficiency of both proteins is required to produce a pro-fibrotic phenotype. Which fibroblasts populations produce Magps and how they limit fibrosis is unknown. We hypothesize that Magp expression is an adaptive response to lung injury and limits fibrosis by sequestering TGFB in ECM and will explore this central hypothesis through three specific aims: 1) Evaluate anti-fibrotic contributions of Magp1 and 2 in pulmonary fibrosis. 2) Evaluate Magp effects on TGFB signaling and immune responses in lung fibrosis. 3) Characterize Magp-producing fibroblast populations. Using combinations of KO and inducible/tissue-specific KO mice, we will determine the individual relationships of these proteins to fibrosis and whether they exhibit redundant antifibrotic functions in vivo. We will analyze alterations in TGFB signaling and immune cell infiltration in lung as possible underlying mechanisms of Magp protection from pulmonary fibrosis. Finally, we will employ our recently developed single nucleus RNASeq approach along with single nucleus assay for transposase-accessible chromatin (snATAC)-Seq to characterize the fibroblast populations that express Magp1 and 2 and evaluate how lung-wide transcriptomes and epigenomes change in the absence of Magps during fibrosis. This proposal will provide Dr. Koenitzer with the mentored training in single cell sequencing and bioinformatics, fibroblast and ECM biology, and lung histology/imaging needed to achieve his goal of independence as a physician-scientist. Under the oversight of an expert scientific advisory committee, He will realize career milestones, complete formal coursework, and develop lasting collaborations. Findings from this work will have implications beyond IPF in other interstitial lung diseases and enable Dr. Koenitzer to develop new approaches to the treatment of lung disease as an independent investigator.

项目摘要/摘要 特发性肺纤维化(IPF)是一种进行性肺部疾病，治疗方案有限，且尚不清楚。 发病机制。增殖的成纤维细胞群体的细胞外基质(ECM)沉积推动了 细胞因子转化生长介导的IPF进行性气流受限和缺氧特性 β因子(TGFb)。直接抑制TGFb会导致无法忍受的副作用，人们对 相反，调整其调节的治疗方法。微原纤维相关糖蛋白1和2(Magp1和Magp2， 或MAGPS)是成纤维细胞产生的蛋白质，分泌到ECM，锚定到微纤维，并结合TGFb 以限制其在基础条件下的信号转导，但在纤维化中没有已知的作用。MAGP基因敲除(KO)小鼠 在各种组织中表现出TGFb信号的增加，并因骨髓而导致免疫细胞改变 TGFb效应。在单细胞RNAseq筛选中，编码这些蛋白质的基因显著上调 在纤维化肺成纤维细胞中，Magp1和Magp2的联合KO导致博莱霉素后更严重的纤维化 对小鼠的损伤。这表明MAGPS作为纤维化信号的抑制因子，并在MAGP1中进行了后续研究 2只KO小鼠表明，缺乏这两种蛋白是产生促纤维化表型所必需的。 目前尚不清楚哪些成纤维细胞群会产生MAGPS，以及它们如何限制纤维化。我们假设 MAGP的表达是对肺损伤的一种适应性反应，并通过隔离ECM和 我将通过三个具体的目标来探讨这一中心假设：1)评估抗纤维化的作用 MAGP1和MAG2在肺纤维化中的表达。2)评估Magp对TGFb信号转导和免疫的影响 肺纤维化的反应。3)鉴定产生MAGP的成纤维细胞群。使用组合 在KO和可诱导/组织特异性KO小鼠中，我们将确定这些蛋白与 纤维化以及它们在体内是否表现出多余的抗纤维化功能。我们将分析TGFb的变化 肺内信号转导和免疫细胞浸润可能是Magp保护肺损伤的机制 肺纤维化。最后，我们将使用我们最近开发的单核RNAseq方法以及 转座酶可及染色质单核测定法(SnATAC-Seq)鉴定成纤维细胞 表达Magp1和Magp2的群体，并评估全肺转录本和表观基因组在 纤维化过程中MAGPS的缺失。该提案将为Koenitzer博士提供指导培训 单细胞测序和生物信息学，成纤维细胞和ECM生物学，以及肺组织学/成像所需的 实现他作为一名内科科学家的独立目标。在专家科学顾问的监督下 作为委员会成员，他将实现职业生涯的里程碑，完成正式课程，并发展持久的合作关系。 这项工作的发现将对IPF以外的其他间质性肺部疾病产生影响，并使Dr。 作为一名独立的研究员，Koenitzer将开发治疗肺部疾病的新方法。